Control of glycogenosis and effects of exercise on phosphorylase kinase and cAMP-dependent protein kinase in rainbow trout organs

Abstract

To analyze the mechanisms of glycogen phosphorylase control in organs of the rainbow trout Oncorhynchus mykiss, activities of glycogen phosphorylase kinase (GPK) and cAMP-dependent protein kinase (PKA), as well as levels of cAMP, were quantified. The complete cascade for activating glycogen phosphorylase was present in trout organs and all components were activated in white skeletal muscle and liver during exhaustive swimming exercise. GPK and PKA showed the highest activities in the liver, being three- and four-fold higher than corresponding activities in white muscle. Exercise stimulated a 60% increase in GPK activity in the liver and a 40% rise in white muscle. Furthermore, the amount of active PKA rose from 12 to 21% in the liver and from 32 to 57% in white muscle after exhaustive exercise and the cellular levels of cAMP increased by 50% in the liver and 70% in white muscle of exercised fish. Other organs (heart, gill, brain, kidney) showed little or no change in these parameters as a result of exhaustive exercise. GPK activity in liver, muscle, and heart extracts was strongly stimulated by in vitro incubation with the catalytic subunit of mammalian PKA, activity rising by 6- to 7-fold in white muscle extracts and 2- to 2.6-fold in liver and heart extracts. This occurred in extracts from both control and exercised fish and suggested that even in fish exercised to exhaustion, the maximal enzymatic potential for activation of glycogenolysis was not expressed. Other modes of GPK activation were not apparent, for the enzyme in crude extracts was stimulated only by incubation with cAMP and did not respond to cGMP or Ca2+ + phorbol 12-myristate 13-acetate. The data indicate that the cAMP-activated, PKA- and GPK-mediated cascade is key to the activation of glycogenolysis in both the skeletal muscle and liver during burst swimming exercise by trout.Key words: exercise, glycogen phosphorylase kinase, protein kinase A, cAMP, Oncorhynchus mykiss, control of glycogenolysis.